This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The N &B technique has been used to study the golgi and plasmamembrane organization of several GPI anchored proteins fused to GFP (Green fluorescent protein) and mCherry, in a MDCK cell line. The analysis has been performed in unpolarized and polarized cells. In order to collect relevant data, cells have been grown respectively in "Matek" chambers and on filters, and image acquisition has been done on live cells, using an Olympus, inverted, laser scanning confocal microscope, equipped with photon counting detector.